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Abstract:

Disclosed is a method for treating or preventing pathological or
non-pathological skin conditions, disorders or diseases comprising
administering a conjugate of proline, hydroxyproline, or a salt thereof,
conjugated with a fatty acid.

Claims:

1. Method for treating or preventing pathological or non-pathological
skin conditions, disorders or diseases comprising administering a
conjugate of proline, hydroxyproline, or a salt thereof, conjugated with
a fatty acid.

3. The method according to claim 1, wherein the conjugate is a compound
of Formula (I) ##STR00005## wherein R1 is selected from hydrogen,
unsaturated mono, poly and omega fatty acids, or OR3; R2 and
R3 are independently selected from hydrogen or unsaturated mono,
poly and omega fatty acids, provided that both R1 and R2 are
not hydrogen and that if R1 is OR3, both R2 and R3
and not hydrogen.

4. The method according to claim 3, wherein R1 is OR.sub.3.

5. The method according to claim 1, wherein the conjugate is a conjugate
of hydroxyproline and docosahexaenoic acid (DHA) according to formula
MW-001: ##STR00006##

6. The method according to claim 1, wherein the skin condition is skin
aging.

8. The method according to claim 1, wherein the conjugate is
co-administered with an antibiotic.

9. The method according to claim 1, wherein a topical, dermatological or
cosmetic formulation comprising said conjugate is administered.

10. The method according to claim 1, wherein an oral formulation
comprising said conjugate is administered.

11. A method for reducing wrinkles comprising the step of administering a
conjugate of proline, hydroxyproline, or a salt thereof, conjugated with
a fatty acid.

Description:

FIELD OF THE INVENTION

[0001] The invention is directed to methods of treating or preventing
pathological and non-pathological skin conditions, disorders and
diseases. The invention is further directed to the treatment of dry skin,
aging skin, hyper-pigmentation, wrinkles as well as acne, irritant diaper
rash, irritant dermatitis, psoriasis, rosacea, impetigo, burns and
wounds.

BACKGROUND OF THE INVENTION

[0002] The L form of proline (2-pyrrolidine-carboxylic acid) and
4-hydroxyproline (4-hydroxy 2-pyrrolidine-carboxylic acid, related to
herein as hydroxyproline) are naturally occurring amino acids and are
found, for example, in relatively large quantities in collagen. Elastins
are also known to contain proline, in about each ninth residue. Proline
and 4-hydroxyproline are not considered to be essential amino acids and
further, no pharmacological activity for non-derivative proline and
4-hydrxyproline is known in the field.

[0003] However, certain N-substituted derivatives of proline or
hydroxproline containing substituted peptides are known ACE-inhibitors
and have become established for their ability to control high blood
pressure (for example Captopril; Lisinopril; Enalapril and Moveltipril).
Oxaceprol, an additionally N-acyl derivative; possesses antiphlogistic;
anti-rheumatic and wound healing activity. Commercial infusions for
parenteral nutrition occasionally contain proline as an adjuvant (for
example, in combination with other amino acids, carbohydrates and
electrolytes).

[0004] Certain Omega-3 and Omega-6 fatty acids are considered essential
fatty acids, which, although essential for human health, cannot be
produced by the human body. Other omega fatty acids are known to be
conditionally essential, i.e., essential to the human body under certain
conditions. Omega fatty acids can be found in fish, seafood and certain
plants. Also known as polyunsaturated fatty acids (PUFAs), omega fatty
acids play a crucial role in brain function, as well as normal growth and
development. They have also become popular since they may reduce the risk
of heart disease. Research shows that omega fatty acids reduce
inflammation and may help lower risk of chronic diseases such as heart
disease, cancer, and arthritis. Omega fatty acids are highly concentrated
in the brain and appear to be important for cognitive and behavioral
function. Symptoms of omega fatty acid deficiency include fatigue, poor
memory, dry skin and heart problems.

[0005] Acne vulgaris is the most common disorder of human skin and is
known to affects up to 80% of the population. Acne has many different
symptoms including comedones, papules, pustules, nodules, cysts and
pilosebaceous inflammation. Among these, inflammatory lesions of acne are
of the greatest concern to patients because they may lead to acne
scarring, thereby inducing adverse psychological effects.
Propionibacterium acnes (referred to herein as P. acnes) is a
Gram-positive anaerobic bacterium that mostly resides in the
pilosebaceous follicles of the skin. Although P. acnes is a member of the
normal skin commensal bacterial flora, it plays a critical role in the
development of inflammatory acne when it overgrows and colonizes the
pilosebaceous unit. It has also been widely accepted that inflammatory
acne is induced by host immune reactions to P. acnes. P. acnes releases
chemoactive factors that attract the immune system cells such as
neutrophils, monocytes, and lymphocytes. Previous studies have found that
P. acnes stimulates the production of proinflammatory cytokines such as
interleukins-1b, -8, -12, and tumor necrosis factor-a. As reduction in P.
acnes numbers in the hair follicle by antimicrobial agents correlates
with clinical improvement of acne in patients, antibiotics have been used
to treat acne for several decades and are still widely prescribed for
acne patients.

[0006] Topical antimicrobial agents include benzoyl peroxide, clindamycin,
erythromycin, tetracycline, azelaic acid, triclosan and various
combinations of these agents. In cases where topical treatment is not
successful or in patients at risk for scarring of the skin and pigmentary
changes, systemic antibiotics are indicated. These include tetracycline,
doxycycline, minocycline and erythromycin. However, data from several
studies indicate a drastic increase in the proportion of patients
carrying P. acnes strains resistant to one or more antibiotics. The
relationship between skin colonization with resistant P. acnes isolates
and treatment outcome is complex and the clinical significance of
resistance is not always entirely clear. Nevertheless, careful analysis
of clinical trials performed over the last two decades shows that there
has been a gradual decrease in the efficacy of topical erythromycin, most
likely related to the development of resistance. It is well established
that bacterial biofilms play an important role in the pathogenesis of
many human infections. One of the striking properties of sessile cells
(i.e. cells growing in a biofilm) is their increased resistance to
antimicrobial agents. Factors considered to be responsible for this
phenotype include restricted penetration of antimicrobials, decreased
growth rate, expression of resistance genes and the presence of resistant
"persister" cells.

SUMMARY OF THE INVENTION

[0007] Embodiments of the invention are directed to a method for treating
or preventing pathological or non-pathological skin conditions, disorders
or diseases comprising administering a conjugate of proline,
hydroxyproline, or a salt thereof, conjugated with a fatty acid.
According to some embodiments, the fatty acid is omega-3 fatty acid,
omega-6 fatty acid or omega-9 fatty acid.

[0008] According to further embodiments, the conjugate is a compound of
Formula (I)

[0009] R2 and R3 are independently selected from hydrogen or
unsaturated mono, poly and omega fatty acids, provided that both R1
and R2 are not hydrogen and that if R1 is OR3, both
R2 and R3 and not hydrogen.

[0010] According to further embodiments, the conjugate is a conjugate of
hydroxyproline and docosahexaenoic acid (DHA) according to formula
MW-001:

##STR00002##

BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The invention is herein described, by way of example only, with
reference to the accompanying drawings. With specific reference now to
the drawings in detail, it is stressed that the particulars shown are by
way of example and for purposes of illustrative discussion of the
preferred embodiments of the present invention only, and are presented in
the cause of providing what is believed to be the most useful and readily
understood description of the principles and conceptual aspects of the
invention. In this regard, no attempt is made to show structural details
of the invention in more detail than is necessary for a fundamental
understanding of the invention, the description taken with the drawings
making apparent to those skilled in the art how the several forms of the
invention may be embodied in practice.

[0012] FIG. 1 is a bar graph presenting the change in the infected area
before and after one and two weeks of treatment.

[0013] FIG. 2 is a bar graph presenting the change in the dryness level
before and after one and two weeks of treatment.

[0014] FIG. 3 is a bar graph presenting the change in the scale level
before and after one and two weeks of treatment.

[0015] FIG. 4 is a bar graph presenting the change in the redness level
before and after one and two weeks of treatment.

[0016] FIG. 5 is a bar graph presenting the change in the lichenification
level before and after one and two weeks of treatment.

[0017] FIG. 6 is a graph presenting the wrinkles appearance before, after
two weeks and after four weeks of treatment.

[0018] FIG. 7 presents the volunteers assessment of wrinkles appearance
after two and four weeks of treatment.

[0019] FIG. 8 presents the volunteers assessment of wrinkles condition
around the eyes after two and four weeks of treatment.

[0020] FIG. 9 presents results of a questionnaire in which the volunteers
were asked to their level of agreement with the statements detailed
therein.

[0021] FIG. 10 presents a comparison between wrinkles around the eyes in
the treated area and the control area.

[0022] FIG. 11 presents a comparison between the firmness of the skin in
the treated area and the control area.

DESCRIPTION OF THE DETAILED EMBODIMENTS OF THE INVENTION

[0023] According to one embodiment of the invention, there is provided a
method for treating or preventing pathological and/or non-pathological
skin conditions, disorders and diseases comprising administering an
amino-acid or salts thereof, conjugated with any appropriate fatty acid
or fatty acid derivative.

[0024] According to one embodiment of the invention, there is provided a
method for treating or preventing pathological and non-pathological skin
conditions, disorders and diseases comprising administering proline or
hydroxyproline, or salts thereof, conjugated with any appropriate fatty
acid or fatty acid derivative.

[0025] According to some embodiments, the conjugate is a compound of
formula (I)

##STR00003##

wherein R1 is selected from hydrogen, unsaturated mono, poly and
omega fatty acids, or OR3; R2 and R3 are independently
selected from hydrogen or unsaturated mono, poly and omega fatty acids,
provided that both R1 and R2 are not hydrogen and that if
R1 is OR3, both R2 and R3 and not hydrogen. According
to some embodiments, the omega fatty acid is omega-3 fatty acid, omega-6
fatty acid or omega-9 fatty acid.

[0026] According to some embodiments, the conjugate is a conjugate between
hydroxyproline and an acid selected from group consisting of a lipoic
acid, oleic acid, linoleic acid, arachidonic acid, eicosapentaenoic acid
(EPA) and docosahexaenoic acid (DHA). According to some embodiments, the
acid is DHA.

[0027] According to some embodiments, the conjugate is a conjugate of
hydroxyproline and docosahexaenoic acid (DHA) according to formula
MW-001:

##STR00004##

[0028] According to some embodiments of the invention, there is provided a
method for inhibiting skin aging, including inhibiting or reducing skin
wrinkles, sagging and/or dry skin comprising topically administering the
conjugate of the invention or a topical, dermatological or cosmetic
formulation including the conjugate of the invention. According to some
embodiments, the conjugate of the invention or a topical, dermatological
or cosmetic formulation including the conjugate of the invention improves
skin aesthetics comprising topically administering the conjugate of the
invention or a topical, dermatological or cosmetic formulation including
the conjugate of the invention. According to further embodiments, the
conjugate of the invention or a topical, dermatological or cosmetic
formulation moisturizes the skin, thereby preventing and/or treating dry
skin.

[0029] According to some embodiments, there is provided a method for
reducing wrinkles comprising the step of administering a conjugate of
proline, hydroxyproline, or a salt thereof, conjugated with a fatty acid.

[0030] According to some embodiments, there is provided a method for
treating acne, irritant diaper rash, irritant dermatitis, psoriasis,
rosacea, impetigo, forunculosis, burns and/or wounds, including chronic
wounds comprising topically administering the conjugate of the invention
or a topical, dermatological or cosmetic formulation including the
conjugate of the invention. According to some embodiments, there is
provided a method for treating acne vulgaris, acne variants, acne
conglobata, acne mechanica, acne excoriee des jeunes filles, drug induced
acne and/or occupational acne comprising topically administering the
conjugate of the invention or a topical, dermatological or cosmetic
formulation including the conjugate of the invention. According to some
embodiments, there is provided a method for treating an infectious skin
disease comprising topically administering the conjugate of the invention
or a topical, dermatological or cosmetic formulation including the
conjugate of the invention. According to some embodiments, there is
provided a method for treating conditions caused by Propionibacterium
acnes comprising topically administering the conjugate of the invention
or a topical, dermatological or cosmetic formulation including the
conjugate of the invention. According to further embodiments, there is
provided a method for treating conditions caused by Staphylococcus spp
comprising topically administering the conjugate of the invention or a
topical, dermatological or cosmetic formulation including the conjugate
of the invention.

[0031] According to some embodiments, the conjugate may be co-administered
with any appropriate antibiotic. When administered to a patient together
with an antibiotic, any type of co-administration is included herein. For
example, the conjugate and the antibiotic may be administered in the same
formulation, at the same time in different formulations or each having
its own administration course, being possibly administered at different
times to the same patient for treating the same condition. The antibiotic
may be administered before, after or at the same time as the conjugate.

[0032] According to some embodiments, synergism is achieved when
co-administering the conjugate and an antibiotic. According to some
embodiments, synergism is achieved when co-administering a conjugate of
hydroxyproline and DHA. According to some embodiments, such synergism
allows the administration of relatively low doses of the conjugate and/or
the antibiotic.

[0033] The conjugate may be administered by any appropriate means,
including topically, orally or parenterally. According to some
embodiments, the conjugate is administered topically in the form of a
topical formulation.

[0034] According to some embodiments, the conjugate is administered
together with any appropriate non-toxic pharmaceutical carrier. The
carrier may be solid or liquid. According to some embodiments, the solid
carrier is selected from lactose, corn starch, gelatin, talc, stearic
acid, magnesium stearate, sucrose, agar, pectin, acacia or any
combination thereof. The liquid carrier may be selected from peanut oil,
olive oil, sesame oil, saline solution, water or any combination thereof.

[0035] According to some embodiments, the conjugate is administered
together with any ingredients appropriate for modifying the release of
the active conjugate from the formulation. Time delaying agents, such as
glyceryl monostearate and glyceryl disearate may be used, with or without
the addition of wax to the formulation.

[0036] According to some embodiments, the topical formulation further
comprises one or more of an anti-inflammatory ingredient, anti-itch
ingredient, anti-hystaminic ingredient, skin hydration ingredient, anti
oxidant, anti microbial ingredient, skin barrier function enhancer, and
any combination thereof. According to some embodiments, the topical
formulation further comprises zinc. According to some embodiments, the
topical formulation further comprises zinc oxide. According to further
embodiments, the topical formulation further comprises silver ions.
According to some embodiments, the topical formulation further comprises
an anti fungal agent. According to some embodiments, the topical
formulation further comprises hydrocortisone. According to some
embodiments, the topical formulation further comprises a corticosteroid.
According to some embodiments, the topical formulation further comprises
a macrolide immunosuppressant. According to some embodiments, the topical
formulation further comprises NSAIDS.

[0037] According to some embodiments of the invention, the pH of the
topical formulation is between about 5.0-7.0. According to further
embodiments of the invention, the pH of the topical formulation is
between about 5-5.5. According to further embodiments of the invention,
the pH of the topical formulation is between about 5.5-6.0. According to
further embodiments of the invention, the pH of the topical formulation
is between about 6.0-6.5. According to further embodiments, the pH of the
topical formulation is between about 6.5-7.0. According to further
embodiments of the invention, the pH of the topical formulation is
between about 5.0-6.5.

[0038] The topical skin formulation may be in the form of a gel, an
ointment, pomade, a body and/or face soap, a cream, a cleansing gel, a
lotion, oil-in-water, water-in-oil, water-in-oil-in-water, and
oil-in-water-in-silicone emulsions, an aqueous solution, a paste, a foam,
a salve, an oil, a wash, a conditioner or an aerosol. The formulation may
be applied in any suitable manner, e.g., by hand, spatula, spray or pad.

[0039] The topical formulation may further comprise cosmetic or
dermatological acceptable ingredients or dermatologically acceptable
carriers. According to some embodiments of the invention, the
compositions may include one or more of an oil, a fat, wax, a detergent,
a conditioner, a pH modifier, a preservative, a solvent, a viscosity
modifier, a colorant, a perfume, a dyestuff and the like.

[0040] According to an embodiment of the invention, the compositions of
the invention are administered once a day. According to other
embodiments, the compositions are administered twice a day, three times a
day or more.

[0041] According to an embodiment of the invention, the composition is
administered chronically. According to some embodiments of the invention,
the composition is administered for about 10 days or more, 20 days, 30
days, 60 days, 90, 120 days or more.

[0042] In some embodiments of the invention, the composition further
comprises a cleaning agent or a detergent that are typically anionic,
cationic, non-ionic or amphoteric surfactants. Typical anionic
surfactants are carboxylates, sulfonates, sulfates or phosphates, e.g.
fatty acid soaps, salts of lauryl sulfate and salts of lauryl ether
sulfate. Examples of cationic surfactants are aliphatic mono, di and
polyamines derived from fatty and rosin acids, amine oxides, ethoxylated
alkyl amines and imidazolines. Examples of non-ionic surfactants are
polyoxyethylene surfactants, alkylphenol ethoxylates, carboxylic acid
esters, e.g., mono and diglycerides, polyoxyethylene esters and fatty
acid diethanolamine condensates. Amphoteric surfactants are those
containing combinations of the anionic and cationic groups described
above, particularly those containing both acid carboxyls and basic
nitrogen groups. Typical amphoteric surfactants are imidazolines and
betaines, e.g., lauric and myristic imidazolines and betaines, and
amidopropylbetaines.

[0043] The topical pharmaceutical compositions may also comprise a
suitable emulsifier, which refers to an agent that enhances or
facilitates mixing and suspending oil-in-water or water-in-oil. The
emulsifying agent used herein may consist of a single emulsifying agent
or may be a nonionic, anionic, cationic or amphoteric surfactant or blend
of two or more such surfactants. Such surface-active agents are described
in "McCutcheon's Detergent and Emulsifiers," North American Edition, 1980
Annual published by the McCutcheon Division, MC Publishing Company, 175
Rock Road, Glen Rock, N.J. 07452, USA.

[0045] According to some embodiments, the conjugate is administered,
together with any additional appropriate ingredients as a formulation in
the form of a food, feed, food additive, feed additive, drink or drink
additive. According to some embodiments, the formulation is in the form
of tablets, capsules, powders, troches, soft gelatin capsules, syrup,
liquid suspension or lozenges and may be administered orally. According
to some embodiments, the conjugate is prepared into a parenteral
formulation, such as for subcutaneous intramuscular or intravenous
administration.

[0046] As defined herein, treating and preventing includes fully healing a
condition, partially healing a condition, such that an improvement occurs
and preventing, or at least partially preventing, the occurrence of the
condition.

[0047] The term "preventing" refers to keeping a disease, disorder or
condition from occurring in a subject. In some cases the subject may be
at risk for developing the disease, but has not yet been diagnosed as
having the disease. In some instances, the term "preventing" refers to
preventing the next cycle of the disease from occurring.

[0049] Atopic dermatitis is a pruritic disease of unknown origin that
usually starts in early infancy and is typified by pruritus, eczematous
lesions, xerosis (dry skin), and lichenification of the skin (thickening
of the skin and an increase in skin markings). The disease usually
appears on the face, on the inner side of the elbows, behind the knees,
and on the hands and feet. Atopic dermatitis is associated with other
atopic diseases (eg, asthma, allergic rhinitis, urticaria, acute allergic
reactions to foods and increased immunoglobulin E ([IgE] production) in
many patients. It is a disease with a high rate of morbidity, and its
occurrence appears to be on the increase.

Inclusion Criteria

[0050] Volunteers aged 2-30 years, with Atopic dermatitis.

Exclusion Criteria

[0051] Volunteers with a known allergy to one of the tested materials or
to any of the ingredients.

[0052] Treatment with medication such as anti-inflammatories,
anti-histamines, corticosteroids, systemically or topically applied,
unless stopped 2 weeks prior to the trial in the case of systemic
treatment and 3 days in the case of topical treatment.

[0053] Volunteers in the process of diagnosis or treatment for cancer,
kidney disease or liver disease.

[0054] Pregnant or lactating women.

Test Performance

[0055] Ten volunteers aged 2-30 years participated in the trial. Four
females and six males.

[0056] Volunteers were instructed to apply the product only on one side of
the body, on the worst affected area, while the other side was defined as
the control area and was not treated. The tested material, a body lotion
product, was applied 1-3 times a day for a period of 4 weeks.

[0057] The volunteers were instructed to avoid using any other treatment
during the test period.

[0058] Each volunteer was examined at baseline (T0), after one week (T1)
and after 2 weeks of using the product (T2). The investigator rated the
condition of the skin according to the parameters of the size of the
affected area, dryness, scale, redness, edema, lichenification and
pruritus. Both the treated and the Control areas were documented with
color photographs at T0, T1 and T2.

[0059] The participants were asked to keep a record of the daily
applications in a diary and to note any side effects resulting from the
product application.

Examination Time-Points

[0060] At baseline (T0)

[0061] After one week of treatment (T1)

[0062] After two weeks of treatment (T2)

Measurements

Hydration Level

[0063] Hydration level was evaluated by Comeometer CM 825, Courage &
Khazaka, CK. The probe of the Corneometer was placed on the skin at a
constant pressure and measurements of skin hydration were taken. Skin
hydration was recorded at baseline, after 1 week and after 2 weeks of
treatment. Room temperature and humidity were kept constant.

Erythema Level

[0064] Erythema level was evaluated by Mexameter, Courage & Khazaka, CK.
The probe of the Mexameter was placed on the skin at a constant pressure
and measurements of skin hydration were taken. Skin erythema level was
recorded at baseline, after 1 week and after 2 weeks of treatment.

Trans Epidermal Water Loss

[0065] Transepidermal water loss (TEWL) was measured by Tewameter 300 TM,
Courage & Khazaka, CK. TEWL was recorded at baseline and after one week
and after two weeks of treatment. Room temperature and humidity were kept
constant.

Study Director's Evaluation

[0066] The following parameters were evaluated:

[0067] a. Lesion area

[0068] b. Dryness

[0069] c. Scales

[0070] d. Redness

[0071] e. Edema

[0072] f. Lichenificalion

[0073] The parameters mentioned above were evaluated on a scale of 0 to 5
by the study director.

[0074] 0=None

[0075] 1=Slight

[0076] 2=Moderate

[0077] 3=Severe

[0078] 4=Extreme

[0079] 5=Very Extreme

Subjective Evaluation

[0080] The volunteers were asked to assess the pruritus intensity on a 0-5
scale at T0, T1 and T2.

[0081] 0=None

[0082] 1=Slight

[0083] 2=Moderate

[0084] 3=Severe

[0085] 4=Extreme

[0086] 5=Very Extreme

Side Effects

[0087] In addition to keeping records of side effects, volunteers were
asked to immediately report any side effects, such as dryness, burning
sensation, peeling of the skin, redness, etc. to 1SR.

Statistical Analysis

[0088] A statistical comparison between T0 and the other two time-points
(T1 and T2) and between the treated and the Control areas were made
according to the Wilcoxon signed-ranks test, while p<0.05 indicated a
significant difference

Test Results

[0089] The results of 10 volunteers at each time point are presented in
FIGS. 1-5. All parameters were assessed in a scale of 0-5 where 0=none
and 5=very extreme. The percentage of change was calculated at T1 and T2.

[0093] Treated area: At T0, the dryness level was 4.1. At T1 the dryness
level was 1.8. At T2 the dryness level was 1.1.

[0094] Control area: At T0, the dryness level was 3.1. At T1 the dryness
level was 3.2. At T2 the dryness level was 1.8.

Statistical Analysis

[0095] After 1 week of treatment, the dryness level decreased
significantly at the treated area. After 2 weeks of treatment, the
dryness level of both sides decreased significantly. Results showing the
change in the dryness level are presented in FIG. 2.

[0099] Treated area: At T0, the redness level was 3.9. At T1, the redness
level was 2.4. At T2 the redness level was 2.3.

[0100] Control area: At T0, the redness level was 3.1. At T1, the redness
level was 3.1. At T2 the redness level was 2.1.

Statistical Analysis

[0101] After 1 week of treatment, the redness level decreased
significantly at the treated area. After 2 weeks of treatment, the
redness level of both sides decreased significantly. Results showing the
change in the redness level are presented in FIG. 4.

[0114] Treated area: The erythema level at the treated area was 620.5 at
T0. At T1 the erythema level was 562.7 level and at T2 the erythema level
was 600.2.

[0115] Control area: The erythema level at the control area was 604.5 at
T0, 604.8 at T1, and 600.2 at T2.

Statistical Analysis

[0116] After 1 week of treatment, the erythema level decreased
significantly at the treated side, but then increased again and was not
significantly different from the erythema level at baseline.

[0117] Referring to the use of the cream, volunteers were asked to fill
out a questionnaire expressing their level of agreement with the
statements listed in the questionnaire on a Scale of 1-10, where the
worst score was 1 and an excellent score was 10. The volunteers assessed
the product as follows: long lasting moisture-7.4; pleasantness-8.6; and
total score was 8.7.

Drop-Outs

[0118] None of the participants dropped out from the test.

Side Effects

[0119] None of the participants had side effects due to product use

Summary

[0120] After two weeks of treatment with the product MW-001 (related to
herein also as MWL-001) there was a significant improvement in the
infected area size, dryness, redness, scale, lichenification and pruritus
at the treated side. There was also a significant improvement in redness
and dryness parameters at the control side.

[0121] All the volunteers were very satisfied with the product, and have
very high intentions of purchasing the product in the future.

[0122] The product MW-001 was found to be safe for use and very efficient
in the treatment of atopic dermatitis.

Example 2

The Efficacy of MW-001 in the Treatment of Psoriasis

Introduction

[0123] Psoriasis is a chronic skin disease common in 2-3% of the
population characterized by faster and uncontrolled growth of skin cells
in specific areas as much as 27 times more than in healthy skin cells. As
a result, the skin cells accumulate layer on layer thus creating thick
skin with "plaque" and inflamed areas are covered with silvery thick
white scales. The affected areas in Psoriasis Vulgaris are usually the
elbows, knees, scalp, back and finger nails

Inclusion Criteria

[0124] Volunteers aged 18-65 with psoriasis

[0125] The study director assigned a serial number to each volunteer.

[0126] Volunteers signed an informed consent form prior to the test and
after detailed explanation.

Exclusion Criteria

[0127] Volunteers with a known allergy to one of the tested materials or
their ingredients.

[0128] Treatment with medication such as anti-inflammatories,
antihistamines, corticosteroids, systemically or topically applied,
unless stopped 2 weeks prior to the trial in the case of systemic
treatment and 3 days before in the case of topical treatment.

[0129] Pregnant or lactating women.

Methodology

[0130] Fifteen volunteers aged 29-73 with psoriasis.

[0131] The volunteers signed a consent form prior to the test and after
detailed explanation.

[0132] The volunteers were instructed to use the cream at least once a
day, for 4 weeks.

[0133] Volunteers were instructed to apply the product only on one side of
the body, on the worst affected area at least once a day, for 4 weeks,
while the other side was defined as the control area and was not treated.

[0134] The volunteers were instructed to avoid using any other treatment
during the test period.

[0135] Each volunteer was examined at baseline (T0), after two weeks (T1)
and after 4 weeks of using the product (T2). The study Director rated the
state of the skin according to the parameters of scales severity,
thickness of the plaque, redness and the size of the lesions. Erythema
level was documented at each time point.

[0136] Each volunteer filled out a questionnaire regarding the tested
cream at the end of the trial.

[0137] The participants were asked to keep a record of daily applications
and to note any side effects resulting from product application.

Examination Time-Points

[0138] At baseline (T0)

[0139] After two weeks of treatment (T1)

[0140] After four weeks of treatment (T2)

Measurements

Erythema Level

[0141] Erythema level was evaluated by Mexameter, Courage & Khazaka, CK.
The probe of the Mexameter was placed on the skin at a constant pressure
find measurements of skin hydration were taken. Skin erythema level was
recorded at baseline, after 2 weeks and after 4 weeks of treatment.

Study Director's Evaluation

[0142] The following parameters were evaluated:

a) Size of lesion b) Thickness of lesion

c) Redness

d) Scales

[0143] The parameters mentioned above were evaluated on a scale of 0 to 5
by the study director.

0=Absent

1=Slight

2=Moderate

3--Severe

4=Extreme

5--Very Extreme

Subjective Evaluation

[0144] The volunteers were asked to assess the pruritus level on a 0-5
scale at T0, T1 and T2.

0=Absent

1=Slight

2=Moderate

3=Severe

4=Extreme

5=Very Extreme

Side Effects

[0145] Volunteers were asked to keep a record and report any side effects,
such as dryness, burning sensation, peeling of the skin, redness or any
other visible changes, and report immediately to study director.

Statistical Analysis

[0146] A statistical comparison between T0 and the other two time-points
(T1 and T2) and between the treated and the Control areas were made by
paired sample t-test, while p<0.05 indicated a significant difference

Test Results

[0147] The results of 15 volunteers at each time point are presented in
the Tables below and in the relevant Figures. All parameters were
assessed in a scale of 0-5 where 0=none and 5=very extreme

Infected Area Size

TABLE-US-00014

[0148] TABLE XVI
T0 T1 T2
Treatment 4.1 3.8 3.9
Control 3.4 3.5 3.3

Treated Area:

[0149] At T0 the infected area size assessment was 4.1. At T1 the infected
area size assessment was 3.8. At T2 the infected area size was 3.9.

Control Area:

[0150] At T0 the infected area size assessment was 3.4. At T1 the infected
area size assessment was 3.5. At T2 the infected area size was 3.3.

Statistical Analysis

[0151] A significant decrease in lesion area size was observed in the
treatment area after 2 weeks of application.

Thickness of the Skin

TABLE-US-00015

[0152] TABLE XV
T0 T1 T2
Treatment 3.0 2.6 2.4
Control 2.9 3.3 3.0

Treated Area:

[0153] At T0 the thickness level was 3.0. At T1 the thickness level was
2.6. At T2 the infected area size was 2.4.

Control Area:

[0154] At T0 the thickness level was 2.9. At T1 the thickness level was
3.3. At T2 the infected area size was 3.0.

Statistical Analysis

[0155] A significant improvement in thickness parameter was observed in
the treatment area after 4 weeks (p<0.05).

Redness Level

TABLE-US-00016

[0156] TABLE XVI
T0 T1 T2
Treatment 3.3 3.5 3.2
Control 3.3 3.1 3.4

Treated Area:

[0157] At T0 the redness level was 3.3. At T1 the redness level was 3.5.
At T2 the redness level was 3.2.

Control Area:

[0158] At T0 the redness level was 3.3. At T1 the redness level was 3.1.
At T2 the redness level was 3.4.

Statistical Analysis

[0159] No significant change in redness parameter was observed in any of
the sides during the whole study period (p>0.05).

Scale Level

TABLE-US-00017

[0160] TABLE XVII
T0 T1 T2
Treatment 4.5 4.1 3.8
Control 4.4 4.6 4.8

[0161] Treated area: At T0 the scales level was 4.5. At T1 the scale
level, assessment was 4.1. At T2 the scale level was 3.8.

[0162] Control area: At T0 the scales level was 4.4. At T1 the scale level
was 4.6. At T2 the scale level was 4.8.

Statistical Analysis

[0163] A significant improvement in scales parameter was observed in the
treatment area after 4 weeks (p<0.05)

Pruritus Level

TABLE-US-00018

[0164] TABLE XVIII
T0 T1 T2
Treatment 2.1 1.6 1.3
Control 1.9 1.6 1.9

[0165] Treated area: At T0 the pruritus level was 2.1. At T1, the pruritus
level was 1.6. At T2 the pruritus level was 1.3.

[0166] Control area: At T0 the pruritus level was 1.9. At T1 the pruritus
level was 1.6. At T2 the pruritus level was 1.9.

Statistical Analysis

[0167] No significant change in pruritus level was observed in any of the
sides during the whole study period (p>0.05)

[0169] Treated area: The erythema level at the treated area was 562.1 at
T0, 517.5 at T1 and 576.7 at T2.

[0170] Control area: The erythema level at the control area was 537.3 at
T0, 478.4 at T1, and 529.1 at T2.

Statistical Analysis

[0171] After 2 weeks of treatment, a significant decrease in erythema
level was observed in the control side (p<0.05)

[0172] Volunteers were asked to answer a questionnaire relating to the use
of the cream and to assess the different qualities of the product and
grade the product in a scale of 1-10, where 1 was the worst score, and 10
was the best score. The volunteers assessed the product as follows: ease
of application-9.1; scent-8.4; spreading on the skin-8.2; long lasting
hydration-5.6; pleasant sensation-6.7; texture-6.5; and total score was
7.2.

Side Effects

[0173] No side effects were observed in this study

Summary

[0174] The general score given to this product was 7.2 in a scale of 1-10.

[0175] The highest scores were for the parameters of scent (8.4) and ease
of application (9.1)

[0176] A significant improvement in the parameters of scales and thickness
of the lesion were observed after 4 weeks of application only at the
treated side.

[0177] After 2 weeks of application, there was a significant decrease in
the lesion area size, but after 4 weeks of application, the decrease was
found to be insignificant.

[0178] Significant differences between the treatment side and the control
side in scale and thickness parameters were observed after both 2 weeks
(T1) and 4 weeks (T2) of application.

[0179] Eight out of 14 volunteers answered that the previous product in
use was better than the study product. Four of these volunteers used
steroids and the other 4 used natural products.

[0180] The results of the study indicate that the product MW-001 is
efficient in the treatment of psoriasis and safe for use.

Example 3

Efficacy, Safety and Tolerance Evaluation Test of the MW-001

Objectives

[0181] Evaluation of the anti wrinkle effect of the products MWL-001 after
4 weeks of use by means of photography, objective assessment, and a
questionnaire as well as silicon replica measuring the skin roughness,
and an evaluation of the effect on skin hydration (moisture) and TEWL
(Trans Epidermal Water Loss).

[0182] Number of Subjects: 10

Inclusion Criteria

[0183] Female subjects aged 40 years and over.

[0184] Absence of any visible skin disease that might be confused with a
skin reaction to the test material.

[0185] No known allergies to skin care products.

[0186] Completion of a Medical history form and the understanding and
signing of an Informed Consent form.

[0187] Considered reliable and capable of following directions.

Exclusion Criteria

[0188] Volunteers with a known allergy to one of the tested materials or
to any of the ingredients.

[0189] Treatment with medication such as anti-inflammatories,
anti-histamines, corticosteroids, systemically or topically applied,
unless stopped 2 weeks prior to the trial in the case of systemic
treatment and 3 days in the case of topical treatment.

[0190] Volunteers in the process of diagnosis or treatment for cancer,
kidney disease or liver disease

[0191] Pregnant or lactating women.

Study Design

[0192] Ten healthy female volunteers were included and completed the
study.

[0193] Prior to the study, each of the subjects received a detailed
explanation from the investigator and signed an informed consent.

[0194] A subjective assessment of facial wrinkles condition was performed
by the volunteers at each time point.

[0195] Color photography was taken at each time point.

[0196] Moisture level of the skin (corneometry) and TEWL (Trans Epidermal
Water Loss) were measured at each time point.

[0197] Silicon imprints were taken from the crow's feet area around the
eyes at both treated and control sides at each time point.

[0198] A cosmetic anti wrinkle product MW-001 was applied on half of the
face for four weeks by each volunteer while the other half served as a
control, without any product application during the whole test period.

[0199] After 2 and 4 weeks of the same parameters of corneometry, TEWL,
color photography, silicone imprints and subjective assessment by the
volunteers of wrinkles condition of both treated and control sides.

[0200] The volunteers filled out a questionnaire after 2 and 4 weeks of
application.

Examination Time-Points

[0201] a. At baseline (T0) b. After 2 weeks of use (T1) c. After 4 weeks
of use (T2)

Methodology

Measurements

Hydration Level

[0202] Hydration level was evaluated by Corneometer, Courage & Khazaka,
CK. The probe of the corneometer was placed on the skin at a constant
pressure and measurements of skin hydration were taken. Skin hydration
was recorded at T0, T1 and T2. Each measurement was repeated three times,
and the average of the repeated measurements was calculated and assembled
in the test results. The room temperature and humidity were kept
constant.

TEWL

[0203] TEWL (the ability not to harm the skin barrier) was measured with
Tewameter TM 300, Courage & Khazaka, CK. TEWL was recorded at T0, T1 and
T2. The results of TEWL in each measurement were recorded after
stabilization of the curve of the results in real time. Room temperature
and humidity were constant.

Silicone Imprints Assessment

[0204] A computer program draws a realistic density three-dimensional
image of the relief of the skin on a colour screen for a particular
density of points selected for x and y axes and then the measurements
data stored in the computer are processed and then evaluated.

[0205] This analysis consists of the following steps:

1. Generation of a roughness profile from the surface relief by alignment
and filtering. 2. Calculation of standardized roughness parameters DIN
4768 ff). The silicone imprints were taken only at the treated side.

Photography

[0206] Color photography of the treated area was taken at T0 (baseline),
T1 (after 2 weeks of treatment) and T2 (after 4 weeks of treatment), in
order to record the wrinkles' appearance.

Subjective Evaluation (Questionnaire)

[0207] The volunteers were asked to answer a questionnaire after four
weeks of application, regarding the assessment of each of the tested
products, and to evaluate the improvement of the skin beneath and at the
side of the eye.

Side Effects/Adverse Events

[0208] Volunteers were asked to report side effects, such as dryness,
burning sensation, peeling of the skin, redness, etc. immediately to the
study director.

Statistical Analysis

[0209] A statistical comparison between the results at T0 and at T1 was
made according to the Wilcoxon signed-ranks test, a very sensitive
paired-samples non-parametric test for small groups with unknown
distribution, where p<0.05 indicates a significant difference. The
software used was SPSS 14.

Corneometry for Moisture Measurements

[0210] The corneometry results are provided in arbitrary units.

[0211] The average hydration level in the treated area was 60.9 at T0,
53.4 at T1 and 53.5 at T2.

[0212] The average hydration level in the control area was 61.7 at T0,
57.6 at T1 and 50.8 at T2.

TEWL--Statistical Analysis

[0213] The average TEWL level in the treated area was 7.8 g/hm2 at
T0, 9.4 g/hm2 at T1 and 7.6 g/hm2 at T2.

[0214] The average TEWL level in the Control area was 6.2 g/hm2 at
T0, 8.8 g/hm2 at T1 and 5.9 g/hm2 at T2.

Wrinkles Appearance with Silicone Imprints

[0215] A graph presenting results of wrinkles appearance with silicone
imprints is presented in FIG. 6.

[0217] Wrinkle's appearance with silicone imprints of the treated
area, compared before and after 2 weeks of use, showed the following
results: Decrease of skin roughness by 10.8% at the treated area.

[0218]
Wrinkle's appearance with silicone imprints of the treated area, compared
before and after 4 weeks of use, showed the following results: Decrease
of skin roughness by 19.5% at the treated area.

[0219] The improvement at
both T1 and T2 was significant (p<0.05)

[0220] Wrinkle appearance by silicone Imprints analysis at the control
area:

[0221] Wrinkle's appearance with silicone imprints of the control
area, compared before and after 2 weeks of use, showed the following
results: Decrease of skin roughness by 0.9% at the Control area.

[0222]
Wrinkle's appearance with silicone imprints of the control area, compared
before and after 4 weeks of use, showed the following results: Decrease
of skin roughness by 1.2% at the Control area.

[0223] The improvement at
both T1 and T2 was found to be not significant (p>0.05)

Questionnaire Results:

[0224] The volunteers were asked to assess the improvement in wrinkles
appearance of both the treated and the Control sides at T1 and T2 in a
scale of 0-10, where 0=no change and 10=very significant improvement. The
results are presented in FIG. 7.

[0225] After 2 weeks of treatment: The volunteers rated the improvement of
wrinkle appearance at the treated side with a grading of 3.5, and the
control area with a grading of 1.6.

[0226] After 2 weeks of treatment: The volunteers rated the improvement in
wrinkles appearance at the treated side with a grading of 4.6, and the
control area with a grading of 1.9.

[0227] The volunteers were asked to assess the improvement of wrinkle
condition around the eyes at both the treated and the control areas at T1
and T2 in a scale of 0-10, where 0=no change and 10=very significant
improvement. The results are presented in FIG. 8.

[0228] After 2 weeks of treatment: The volunteers rated wrinkle
improvement at the treated side with a grading of 3.5 and the control
area with a grading of 1.3.

[0229] After 4 weeks of treatment: The volunteers rated wrinkle
improvement at the treated side with a grading of 4.3, and the control
area with a grading of 1.7.

[0230] FIG. 9 presents results of a questionnaire in which the volunteers
were asked to their level of agreement with the statements appearing on
the questionnaire, wherein the scale was from 1-4 (1=total disagreement,
4=total agreement).

[0231] FIGS. 10 and 11 present a comparison between the treated area and
the control area in the parameters of wrinkles around the eyes and
firmness of the skin.

Side Effects/Adverse Events

[0232] None of the volunteers reported of any side effects.

CONCLUSIONS

[0233] On a test panel of 10 subjects who were included and completed the
test after 4 weeks of use, the following conclusions were reached:

[0234] There was a significant improvement in wrinkles' condition around
the eye at the treated side according to the analysis of the silicone
imprints. The control area showed no significant change.

[0235] All the
tested parameters that were assessed by the volunteers showed better
results at the treated area when compared to the control area.

[0236]
Seven out of 10 participants intend to purchase the product in the
future.

[0237] No side effects were observed.

[0238] These results indicate that the product MW-001 is efficient for the
treatment of rough and wrinkled skin, and is safe for use.